This investigation examined the effects of zinc status on cell proliferation and the synergic roles of the metallothionein (MT) and zinc transporter (ZnT) in the human colon adenocarcinoma cell line Caco-2. Cells were treated with 0 to 300 micromol/L ZnSO(4) or 0 to 10 micromol/L N,N,N',N'-tetrakis(2-phridylmethyl) ethylenediamine (TPEN). Cell proliferation was determined by MTT assay and apoptotic cells detected by flow cytometry (Hoechst 33258 dye). mRNA expression of MT1; ZnT1; zrt, irt-like protein 1, 4 (ZIP1, 4); and divalent metal transporter (DMT1) were determined by the reverse transcription polymerase chain reaction or real-time polymerase chain reaction. The results showed that either high or low zinc could inhibit the cell proliferation. The number of apoptotic cells increased with incremental increases in the concentrations of ZnSO(4) and TPEN. The mRNA expression of ZnT1 and MT1 responded significantly after 6 and 12 hours with 200 micromol/L zinc treatment, respectively, and increased gradually with zinc levels from 0 to 200 micromol/L. Compared with the unchanged ZIP1 mRNA expression, ZIP4 was closely dependent on TPEN treatment duration and concentration. The DMT1 mRNA expression was upregulated time-dependently but not concentration-dependently in the late TPEN treatment duration. The results suggest that ZIP4 and DMT1 mRNA expressions are susceptible to low extracellular zinc concentration and upregulated to enhance zinc absorption, whereas the ZnT1 and MT1 act as the key regulators under high zinc conditions to enhance the intracellular zinc efflux to maintain zinc homeostasis. We propose that in response to variations in zinc concentration, the cooperated regulative roles of ZnT1, MT1, DMT1, and ZIP4 contribute to zinc homeostasis.